Investigation of Wound Healing Properties of Aqueous Extracts of Caucasus Herbs at Diabetes Mellitus

INTRODUCTION

Today, diabetes mellitus is one of the most common pathologies that leads to severe systemic complications [1-3]. High titers of corticosteroids initiate a cascade of catabolic processes, which affect the deactivation of regeneration processes of both epithelial tissues and wound processes [4-6]. The corrective factor of dysmetabolic disorders caused by diabetes mellitus is a directed diet [7, 8]. In this case, the greatest positive effect is achieved by combining pharmacological and nutritional components (hypocholesterolemic, antioxidant, hypoglycemic, and immunomodulatory effects) [9-11].

Phytotherapy is a recognized treatment method in official and folk medicine [12-14]. The interest in this method of exposure is increasing due to its lower toxicity and greater commitment to use, especially in countries with traditionally developed folk medicine [15, 16]. Cymbopogon proximus, Acacia nilotica, Trigonella foenumgraecum, and Suregis rotundus have a wide range of pharmacological effects [17, 18]. Among them, the most valuable are antiatherogenic, antidiabetic, antianorexic, antioxidant, anti-carcinogenic, hypolipidemic, and anti-inflammatory effects [19-22].

It is worth noting that in patients with type 2 diabetes mellitus, as well as mild diabetes mellitus, phytotherapy can be used in the form of monotherapy, providing stabilization or regression of the disease [23, 24]. Prolonged use of herbal remedies for this disease improves the general well-being of patients, and their quality of life, in general, reduces hyperglycemia, which allows you to reduce doses of antidiabetic drugs and even sometimes do without them in the future [25, 26].

Therefore, this work aimed to study the regenerative processes of an infected wound in hyperglycemia using dosage forms based on Caucasus herbs: Acacia nilotica, Cyperus rotundus, Trigonella foenumgraecum, and Cymbopogon proximus.

MATERIALS AND METHODS

The study was performed on 35 nonlinear white rats of both sexes weighing 200-250 g. All animal manipulations were carried out by the Guidelines for the Maintenance and Use of Laboratory Animals in the Russian Federation.

Modeling of hyperglycemia was achieved by intramuscular administration of 0.2 mL of 4% dexamethasone solution for 4 days. The skin on the back was treated with depilatory cream, and cleaned, local anesthesia with 0.25% novocaine solution was previously performed. Then, a 2 cm razor incision (0.5 mm deep) was applied to the prepared zone [27]. A double application of Streptococcus epidermidis-infected material from a patient with streptoderma was performed in the wound area.

All experimental animals were randomly divided into 7 groups of 5 rats each: intact group (group 1), 1^st control group where the wound was treated with vaseline (group 2), 2^nd control group where the wound was treated with fusidine (group 3), Experimental group where the wound was treated with an emulsion containing C. proximus and rats received orally an aqueous extract of C. proximus (group 4), an experimental group with topical application of ointment based on C. rotundus and vaseline and oral administration of aqueous extract of C. rotundus (group 5), experimental group with oral administration of aqueous extract of A. nilotica and topical application of an ointment based on A. nilotica (group 6), an experimental group with oral administration of an aqueous extract of Tr. foenumgraecum and topical application of ointment based on Tr. foenumgraecum (group 7).

On the 5^th day of the experiment, a visual assessment of local changes in the wound area was performed according to the following semi-quantitative criteria (on the plus side – compaction, roughness, pain on palpation, hyperemia, swelling, regeneration) [28, 29]. The severity of the signs was assessed on a scale where the maximum severity of the signs was estimated at 5 points, the absence of a sign – at 0 points [30, 31].

Blood was taken from the caudal vein, and stabilized with a 10% sodium citrate solution, the blood composition and biochemical composition of the blood were determined using analyzers PCE-90Vet (USA) and HUMASTAR 600 (Germany), respectively [32, 33]. On the 5^th day, the animals were removed from the experiment in the chamber with ether for anesthesia.

Statistical data processing was performed using the STATISTICA 12 software. The critical level of significance in testing statistical hypotheses in this study was assumed to be 0.05. In the course of the presentation, statistical indicators are indicated by the following symbols: M is the sample average, m (SEM) is the error of the average, and p is the achieved level of significance at the value.

RESULTS AND DISCUSSION

It was found that the administration of dexamethasone solution to rats at the rate of 800 µg/kg for 4 days leads to the development of hyperglycemia (Figure 1).

An increase in target glucose titers was recorded 6 times – up to 30.02 ± 5.16 mmol/L with an initial level of 5.08 ± 1.18 mmol/L, p<0.001. Oral administration of aqueous extracts from the 1^st day of dexamethasone administration helped to correct glucose levels. The maximum hypoglycemic effect was recorded with oral administration of extracts of Acacia nilotica and Cyperus rotundus, showing that hypoglycemia occurred only 2.74 times, p<0.001. Oral administration of Trigonella foenumgraecum extracts showed an increase in glycemia by 304%, and Cymbopogon proximus by 329%, p<0.001.

After discontinuation of dexamethasone solution, the glucose titer in the group of animals without correction decreased by 65% and amounted to 10.4 ± 2.85 mmol/L, p<0.001, but remained almost 2 times higher than in intact animals, p<0.001. Persistent hyperglycemia persisted after dexamethasone administration. Oral administration of aqueous extracts containing the studied herbs and topical application of ointments had a pronounced hypoglycemic effect. This was most pronounced against the background of the introduction of Acacia nilotica and Cyperus rotundus. Trigonella foenumgraecum and Cymbopogon proximus reduced glucose level to 6.32 ± 0.58 mmol/L and 5.4 ± 0.79 mmol/L, respectively, p<0.01. Interestingly, fusidine was found to increase the activity of hepatic transaminases: ALT was increased by 341% and AST by 134%, p<0.01 (Table 1).

Note: * – statistical significance of the difference about the indicators of the group of intact animals at p<0.05; # – statistical significance of the difference in relation to the indicators of the group of animals treated with fusidine at p<0.05; +* – statistical significance of the difference in relation to the indicators of the control group at p<0.05.

The studied dosage forms containing Trigonella foenumgraecum, Cymbopogon proximus, and Acacia nilotica plants prevent the growth of ALT by 34, 35.5, and 38.6%, respectively, p<0.001. Against the background of the use of Cyperus rotundus dosage forms, a decrease in AST activity by 59% is observed, and a statistically significant increase in the level of nitrogenous slags is also observed against the background of fusidine (+18.2%), Trigonella foenumgraecum (+118%), Cyperus rotundus (+137%), p<0.001. It is worth noting that an increase in urea levels was not accompanied by an increase in creatinine, which, of course, indicates not a violation of kidney function, but the induction of protein-synthetic processes and an increase in urea-synthesizing liver function [34, 35].

Only against the background of oral administration of Acacia nilotica aqueous extract, there was a statistically significant increase in albumins by 14%, p<0.001. Against the background of the use of Trigonella foenumgraecum aqueous extract, there was a statistically significant increase in total cholesterol by 49%, p<0.001. Notably, the positive dynamics of metabolic processes and metabolic shifts can be considered as a trigger component of stimulation of regenerative and reparative mechanisms, against the background of phytotherapy [36, 37].

In the general analysis of peripheral blood in animals in the control series using fusidine, leuko-, lymph- and monocytopenia, as well as granulocytosis, were detected. The number of red blood cells increased by 93%, hemoglobin increased by 148%, and a decrease in the number of platelets by 38.8% was also revealed in comparison with the indicators in a series of animals treated with vaseline, p<0.001 (Tables 2 and 3).

Note: * – statistical significance of the difference in relation to the indicators of the group of intact animals at p<0.01; # – statistical significance in relation to the indicators of the control group (vaseline) at p<0.01.

Dosage forms containing Cyperus rotundus and Cymbopogon proximus prevent the development of leukopenia in control animals treated with fusidine (an increase in the number of leukocytes by 95% and 215%, respectively). Dosage forms containing Trigonella foenumgraecum and Acacia nilotica did not affect the correction of leukopenia.

Note: * – statistical significance of the difference in relation to the indicators of the group of intact animals at p<0.01; # – statistical significance of the difference in relation to the indicators of the group of animals treated with fusidine at p<0.01; +* – statistical significance of the difference in relation to the indicators of the control group (vaseline) at p<0.01.

Emulsions containing Cyperus rotundus and Cymbopogon proximus lead to lymphocyte growth by 484% and 383%, respectively, but do not correct granulocytopenia and monocytopenia. Acacia nilotica, Trigonella foenumgraecum, and Cyperus rotundus cause an increase in platelet levels by 3.76, 3.2, and 6.72 times, respectively, p<0.01. The use of dosage forms containing Cymbopogon proximus, Acacia nilotica, Trigonella foenumgraecum, and Cyperus rotundus contributed to maintaining a high level of erythrocytes, which was comparable to those in the fusidine series (control), statistically significantly higher than in the vaseline series, p<0.01 (Table 4).

Note: * – statistical significance of the difference in relation to the indicators of the group of intact animals at p<0.01; # – statistical significance of the difference in relation to the indicators of the group of animals treated with fusidine at p<0.01; +* – statistical significance of the difference in relation to the indicators of the control group (vaseline) at p<0.01.